Process for separating and deprotecting oligonucleotides

C - Chemistry – Metallurgy – 07 – H

Patent

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C07H 1/06 (2006.01) C07H 21/00 (2006.01)

Patent

CA 2494150

A method for deprotecting oligonucleotides comprising: (a) providing protected oligonucleotides wherein (i) the 5' end is linked to a hydrophobic separation function (e.g. dimethoxytrityl) or (ii) both ends are linked to a pair of hydrophobic separation functions in which pair one member is less hydrophobic than the other one (e.g. being alkyl, aryalkyl or arylsiloxyl); (b) precipitating (e.g. by drying) the oligonucleotides on a hydrophobic support (e.g. a hydrophobic polystyrene-based support) using an organic solvent, thereby non-covalently immobilizing the oligonucleotides and making them insoluble; and (c) deprotecting (i.e. removing the hydrophobic separation functions) the oligonucleotides using a reagent in an organic solvent (e.g. 2 % trichloroacetic acid in dichloromethane).

L'invention concerne un procédé de déprotection d'oligonucléotides, consistant :(a) à fournir des oligonucléotides protégés dans lesquels (i) l'extrémité 5' est reliée à une fonction de séparation hydrophobe (du diméthoxytrityle, par exemple) ou (ii) les deux extrémités sont reliées à une paire de fonctions de séparation hydrophobes dont un élément est moins hydrophobe que l'autre (un alkyle, un arylalkyle ou un arylsiloxyle, par exemple) ; (b) à précipiter (par séchage, par exemple) les oligonucléotides sur un support hydrophobe (un support à base de polystyrène hydrophobe, par exemple) à l'aide d'un solvant organique, ce qui a pour effet d'immobiliser les oligonucléotides de façon non covalente et de les rendre insolubles ; et (c) à déprotéger (c'est-à-dire à retirer les fonctions de séparation hydrophobes) les oligonucléotides en utilisant un réactif dans un solvant organique (2 % d'acide trichloroacétique dans du dichlorométhane, par exemple).

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